Supplement foods and pharmaceuticals

ABSTRACT

The present invention provides effective supplement foods and safe pharmaceuticals, each having a hypotensive effect and anti-diabetic effect and no adverse effects. The supplement foods and pharmaceuticals of the present invention comprise squash extract mixed with onion extract. Synergistic effect of the two extracts can be expected by mixing the onion extract with the squash extract.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a divisional of U.S. patent application Ser. No.10/978,718 filed on Nov. 1, 2004, the disclosure of which is herebyincorporated by reference in its entirety, which claims priority under35 U.S.C. §119 to Japanese Patent Application Serial No. JP2004-58837,filed on Mar. 3, 2004.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to supplement foods and pharmaceuticalscomposed of squash extract mixed with onion extract.

2. Description of the Related Art

It has been conventionally known that onion extract has an anti-diabeticeffect (see Japanese Patent Laid-Open No. 10-77232); Yoshimi Saitou,Effects of Onion Concentrated Dried Tablets (OCDT) on PostprandialHyperglycemia in Diabetic Cases, Rinsho Iyaku (Clinical Medicines), Vol.17 No. 7, 1089-1095 (2001); Yoshimi Saitou, Effects of OnionConcentrated Dried Tablets (OCDT) on Hyperlipidemia in Diabetic Cases,Rinsho Iyaku (Clinical Medicines), Vol. 17 No. 8, 1216-1221 (2001);Kazuhiko Yoshizawa, Yoshitaka Aiso, Clinical Effect of Tamaekisu on TypeII Diabetics, Japan Diabetic Society Kanto-Koshinetsu District 41stLecture Abstract, p. 36 (2004)). It is also known that onion extract hasa hypotensive effect (see, for example, Yoshimi Saitou, Effects of OnionConcentrated Dried Tablets (OCDT) on Postprandial Hyperglycemia inDiabetic Cases, Rinsho Iyaku (Clinical Medicines), Vol. 17 No. 7,1089-1095 (2001)). The hypotensive effect of squash extract is based onits angiotensin converting enzyme inhibitory effect (ACE inhibitoryeffect). This was discovered by Abe and Kosaka among the inventors, andfiled by the applicants (Japanese Patent Application No. 2002-312459).

Hypotensive agents having an ACE inhibitory effect (for example, calciumantagonists) have actions, which include actions of suppressing the newonset of diabetes, and actions of suppressing the onset of complicationsin diabetic patients (see, for example, Hansson L et al.: Effect ofangiotensin-converting-enzyme inhibition compared with conventionaltherapy on cardiovascular morbidity and mortality in hypertension: theCaptoril Prevention Project (CAPPP) randomized trial, Lancet 353:611,1999; Yusuf S et al.; Effect of angiotensin-converting-enzyme inhibitor,ramipuril, on cardiovascular events in high-risk patients: The HeartOutcome Prevention Evaluation Study Investigators, N. Engl J Med342:145, 2000). Its mechanism of action is to cause an increase insensitivity to insulin, i.e. the reduced action of insulin resistancehas been suggested (see, for example, Tetsuya Shinai, MasatsuguHoriuchi, Angiotensin and Diabetic Complications, Gendai Iyaku (ModernMedicines), Vol. 35 No. 9, (2003)).

It is said that there are approximately 14 million diabetic patientsincluding potential patients, and about 40% are presumed to be receivingmedical drug therapy. However, in the case of medical drug therapy,adverse effects are of concern. The tremendous cost of medical drugtherapy is also a problem in view of medical economics.

It is therefore desirable for patients with mild diabetes or potentialpatients about to develop diabetes to utilize effective supplement food(processed food) etc., if available, without receiving any medical drugtherapy.

However, currently commercially available processed foods of onionextract such as those seen in Japanese Patent Laid-Open No. 10-77232have not been satisfactory with respect to their effect.

The present inventors discovered that squash extract mixed with onionextract has excellent ACE inhibitory activity (described later inExamples 1 to 3), and therefore have filed an application entitled,“Supplementary Foods Effective In Reducing Blood Pressure” (JapanesePatent Application No. 2002-312459). As mentioned above, since thehypotensive effect based on ACE inhibitory activity also suppresses newonset diabetes, it is expected that supplement foods containing squashextract mixed with onion extract will have an anti-diabetic effect aswell as the hypotensive effect. When these supplement foods wereadministered to diabetic patients with hypertension complications,extremely good performance as described later was obtained.

Supplement foods containing squash extract mixed with onion extract maynot only be used as supplement foods (processed foods), but also aspharmaceuticals.

The object of the present invention therefore is to provide moreeffective supplement foods and safe pharmaceuticals with no adverseeffects by means of both the hypotensive effect and anti-diabeticeffect.

SUMMARY OF THE INVENTION

In order to accomplish the objective, the supplement food of the presentinvention comprises squash extract mixed with onion extract.

Further, the pharmaceutical of the present invention comprises squashextract mixed with onion extract.

Both squash and onion have been consumed by human beings for the pastfew thousand years as foodstuff. Their safety can well be presumed fromthis abundant experience in consuming, and they are available as rawmaterial at low costs.

In addition, as they have been consumed for a long time and their safetyis empirically proven as described above, by mixing squash extractpowder with onion extract powder, then adding pharmaceuticallyacceptable additives (for example excipients, surfactants, etc.) asrequired, safe pharmaceuticals without adverse effects can be prepared.

By mixing squash extract with onion extract, a synergistic effect of thetwo extracts is expected, thus providing, for example, supplement foodsand pharmaceuticals with combined hypotensive and anti-diabetic effects,which are safe and more effective than onion extract, etc. alone, andhave no adverse effects.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the changes of systolic and diastolic blood pressure afterthe ingestion of the products mixed with squash extract and onionextract;

FIG. 2 shows the changes of substances related to elevation in bloodpressure and hypotensive minerals after the ingestion of the productsmixed with squash extract and onion extract;

FIG. 3 shows the correlation between systolic blood pressure andsubstances related to elevation in blood pressure; and

FIG. 4 shows the changes of 2 hour postprandial blood glucose levels andHbA_(1c) after the ingestion of the products mixed with squash extractand onion extract.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Preparation examples and samples will now be described.

PREPARATION EXAMPLE 1

Process for Preparing Squash Extract

(Preprocess)

First, 78 kg of squash (cultivar Ebisu) was washed with Jet Washer(Arimitsu Industry Co., Ltd., SKY-130II) to remove surface soil, thenpeeled and deseeded by hand to obtain flesh. The flesh was grinded intoa size of few millimeters with a Feather Mill (Hosokawa MicronCorporation, FM-65042) to obtain 56 kg of ground product.

(Preparation of Squeezed Liquid)

An equivalent amount of boiling water was prepared in a double-bottompot, and then the ground product was added to the pot and the mixturewas left at 90° C. for 10 minutes. The mixture was placed in a bag offilter fabric and squeezed with oil hydraulic press (Komagata KikaiSeisakusho, KS-3) to obtain 90 kg of squeezed liquid. Soluble solidmatter content of this liquid was 5.4%.

(Preparation of Concentrated Liquid)

To reduce viscosity, 0.01% by weight of enzyme (Sankyo Co., Ltd.,Sucrase N) was added to the squeezed liquid, and the mixture was left at45° C. for 1 hour and then heated to 90° C. to inactivate the enzymecompletely. This liquid was vacuum concentrated under conditions of aheating temperature of 120° C., 160 mmHg, using a concentration pot(Yashima Chemical Engineering Co., Ltd., Type AS) to obtain 13 kg ofconcentrated liquid. Soluble solid matter content of this liquid was36%.

(Preparation of Powder)

Food grade dextrin (Nissi Co., Ltd., NSD 500) 14 kg was dissolved in 28kg of hot water, then heated at 95° C. for 20 minutes. The obtainedconcentrated liquid (13 kg) was added and the mixture was stirred tohomogenize at 70° C. until uniform. Then, this was converted into powderusing a spray drier (Ohkawara Kakohki Co., Ltd., Type L-12) underconditions of a blower temperature 150° C. and exhauster temperature 70°C., sieved with 30-mesh to obtain 17.5 kg of squash extract powder.

PREPARATION EXAMPLE 2

Process for Preparing Onion Extract

Ten kg of onions from Hokkaido were washed twice with water, thenimmediately put in a pot and heated to boil at 100° C. The boiled onionsobtained were ground in a feather mill and squeezed with a squeezingapparatus to obtain 9.5 kg of squeezed liquid. Vacuum concentration wascarried out using a heating temperature of 92° C., evaporationtemperature of 42° C., and pressure of 15 mmHg using a centrifugal-filmvacuum concentrator to obtain 0.9 kg of concentrated extract. Theconstituents distilled off during the vacuum concentration were passedthrough an activated charcoal layer to be adsorbed thereon. The adsorbedmaterial was eluted with ethanol, and then the ethanol was removed fromthe solution by evaporation to obtain 1.5 g of residual component. Thiswas added uniformly to the concentrated extract obtained earlier toobtain 0.9 kg of onion extract.

EXAMPLE 1

Results of Study of ACE Inhibitory Activity and Interaction betweenSquash and Onion Extracts

Measurements were carried out according to the following partly modifiedCushman & Cheung's method [Seikagaku Jikkenhou 38: Shokuhinchu noSeitaikinou Chosetsubusshitsu Kenkyuhou (Experimental Methods inBiochemistry 38: Methods of Researching Substances Regulating VitalFunctions in Food Materials), Shunro Kawagishi ed., Gakkai ShuppanCenter (1996)].

[Samples]

Squash extract (Nippon Shinyaku Co. Ltd., Chitose Foodstuffs Plant)

Onion extract (Nippon Shinyaku Co. Ltd., Chitose Foodstuffs Plant)

Commercially available food product A (specified health food containinglactotripeptide)

Commercially available food product B (specified heath food containingsardine peptide)

[Method]

Reagents

Buffer: 450 ml of 50 mM Na₂B₄O₇ and 550 ml of 200 mM H₃BO₃ are mixed andadjusted to pH 8.3.

Substrate solution: 7.6 mM Hip-His-Leu and 608 mM NaCl are dissolved inthis buffer solution.

ACE solution: ACE is dissolved in the above buffer (60 mU/ml ofactivity).

Method of Measurement

30 μl of a sample solution and 250 μl of a substrate solution are placedin a test tube, and then incubated in a thermobath at 37° C. for 5minutes.

100 μl of ACE solution (6 mU) is added, immediately stirred, followed byreaction at 37° C. for 30 minutes.

250 μl of 1 N hydrochloric acid is added and stirred to terminate thereaction. 1.5 ml of ethyl acetate is added, stirred well, followed byextraction of hippurate released.

Following centrifugation at 3000 rpm for 10 minutes, 0.5 ml of an ethylacetate upper layer is recovered and put in a test tube.

Ethyl acetate is aspirated and removed in a desiccator. After confirmingthe complete removal of ethyl acetate, 4 ml of distilled water is addedand stirred well to dissolve the hippurate, and absorbance is measuredat 228 nm.

Inhibition rate is represented by:${{Inhibition}\quad(\%)} = {\frac{{Ec} - {Es}}{{Ec} - {EB}} \times 100}$

Wherein Es is an absorbance when sample solution is added, Ec is anabsorbance when distilled water instead of a sample solution is added,and EB is an absorbance when reacted after adding 1 N hydrochloric acid.

Calculation of Inhibitory Activity

The concentration of the sample in a reaction solution when aninhibition rate is 50% is defined as the IC50 value. Glucose contents ofall samples are adjusted to 2.0° Bx before measurement.

[Results] 1) Single Materials Squash Onion Commercially commerciallyextract extract available food available food Sample (B × 2.0) (B × 2.0)product A product B ACE 23.7 21.1 79.0 28.9 inhibitory activity IC₅₀(μl/ml) 2) Mixture of Equal Amounts Commercially Commercially Squashextract + available food available food onion extract product A +product B + Sample (each B × 2.0) onion extract onion extract ACE 10.540.2 25.0 inhibitory activity IC₅₀ (μl/ml)

[Discussion]

Considerably high ACE inhibitory activity was seen with both of thesquash extract alone and the onion extract alone.

The activities of leading products known to be specified health foods(commercially available food product A and commercially available foodproduce B) are as shown above; they showed lower activities than thesquash extract alone and the onion extract alone.

In the case of squash extract+onion extract, a synergistic effect wasnotably seen; ACE inhibitory activity was prone to increase by 2-fold.

In the cases of commercially available food product A+onion extract andcommercially available food product B+onion extract, no synergisticeffect was seen (additive effect).

As can be considered from comparison to commercially available foodproducts A and B, the amount of ingestion per day for squash extractappears to be approximately 3 to 10 g as powder containing 30% extractsolid matter.

EXAMPLE 2

Clinical Study of Effects of the Products containing Squash ExtractMixed With Onion Extract on Hypotensive and Hypoglycemic Actions inDiabetic Patients With Hypertensive Complications (Yoshimi Saitou, HeadSurgeon, Bunkyo Daiichi Iin)

Object

We have already confirmed the hypotensive effect of products containingsquash extract mixed with onion extract, although its hypoglycemiceffect in diabetic patients has not yet been studied. Hypoglycemiceffect of onion extract alone has already been confirmed, althoughhypoglycemic effect when mixed with extract of squash extract has notyet been studied. The object of the present test is to examine thehypoglycemic effect of the products containing squash extract mixed withonion extract, compared with onion extract alone.

Methods

Subjects were 8 diabetic patients with hypertensive complications (3males, 51, 59, and 70 years old; 5 females, 51, 61, 65, 68, and 70 yearsold). Three patients received hypotensive agents (calcium antagonists)concomitantly and 5 patients took no concomitant drugs. Seven patientsreceived concomitant anti-diabetic drugs and 1 patient received noconcomitant drugs. The 20 pills of products containing squash extractmixed with onion extract were ingested divided into 2 portions, 10 pillseach, every morning and night. The observation period was set at 16weeks, and the patients were followed before the intake and every 4weeks afterwards. Items examined were: Measurement of blood pressure,every 4 weeks (time of measurement was unified to either a.m. or p.m.);measurement of substances related to elevation of blood pressure(aldosterone, ACE, angiotensin II), before the intake and 8 and 16 weeksafter the intake for a total of 3 measurements; measurement ofhypotensive minerals (K, Ca, Mg), before the intake and 8 and 16 weeksafter the intake for a total of 3 measurements; and measurement of bloodglucose and Hb_(A1C), every 4 weeks.

Results

The changes of systolic and diastolic blood pressure are shown inFIG. 1. Systolic and diastolic blood pressure decreased significantly at4 weeks after beginning administration with the products containingsquash extract mixed with onion extract, and subsequently graduallydecreased. When observed case by case, the hypotensive effect was proneto be more significant when the pressure before beginning theadministration was higher. Concomitant use of hypotensive agents did notseem to be correlated with the effect of the present product. Amongsubstances related to elevation of blood pressure, angiotensin 11 andACE were prone to decrease over time, and aldosterone showed virtuallyno change (FIG. 2). Hypotensive minerals Ca, K, and Mg also showedvirtually no change (FIG. 2). As a result of examining correlationbetween systolic blood pressure and the substances related to elevationin blood pressure, there was a positive correlation between systolicblood pressure and ACE. That is, ACE decreased with decrease in bloodpressure (FIG. 3). Similar trend was seen between diastolic bloodpressure and ACE. The changes of postprandial blood glucose level areshown in FIG. 4. There was clearly a decrease in postprandial bloodglucose level after beginning the administration, and statisticallysignificant decrease was seen after 8 weeks. Hb_(A1C) tended todecrease.

Discussion

It is hypothesized that the products mixed with squash extract and onionextract suppress ACE, and as a result suppress the production ofangiotensin II, thus enhancing the hypotensive effect. It should beparticularly noted that better postprandial hypoglycemic effect wasseen, compared to onion extract alone as previously reported by us. Fromthis fact, it can be hypothesized that squash extract also lowers bloodglucose levels. Recently, hypotensive agents (ACE inhibitors) such asCAPTOPRIL® are reported to suppress the onset in diabetic patients andto suppress diabetic complications. The relationship between ACEinhibitor effect and reduced insulin resistance effect is beingconsidered, and the action mechanism of squash extract in terms ofhypoglycemic effect is thought to be due to insulin resistance reducingeffect based on the ACE inhibitory effect of squash extract.

CONCLUSION

The products containing squash extract mixed with onion extract wereproved to have hypotensive effect, as well as hypoglycemic effectstronger than that seen with onion extract alone. This mechanism ofaction was presumed to be from the insulin resistance reduction effectbased on the ACE inhibitory effect of squash.

1. A method of treating high blood pressure and diabetes comprising thestep of administering a food supplement or a pharmaceutical to a subjectcomprising a combination of a squash extract and an onion extract;wherein the squash extract is produced by a process comprising the stepsof: grinding squash flesh; mixing the ground squash flesh with water;squeezing the ground squash flesh with water through a filter to form asqueezed liquid; adding an enzyme to the squeezed liquid; vacuumconcentrating the squeezed liquid to obtain a first concentrated liquid;dissolving a dextrin in water to form a dextrin and water mixture andadding the first concentrated liquid to the dextrin and water mixture toform a second concentrate; and drying the second concentrate; andwherein the onion extract is produced by a process comprising the stepsof: boiling onion to form a boiled onion; grinding the boiled onion;squeezing the ground, boiled onion to obtain a squeezed liquid; andvacuum concentrating the squeezed liquid.
 2. The method of claim 1,wherein a pharmaceutical is administered to the subject and thepharmaceutical further comprises a surfactant.
 3. The method of claim 2,wherein the pharmaceutical further comprises an excipient.
 4. The methodof claim 1, wherein the pharmaceutical further comprises an excipient.